Biomedical Engineering Reference
In-Depth Information
Table 5.2 Examples of cell types and materials successfully deposited by LDW, illustrating the
applicability and broad range of research
(cont.)
Ribbon material (not
including laser-transparent
ribbon or cell suspension)
Receiving substrate
material
Cell type
References
Rat B35 neuronal
neuroblast cells
Matrigel
®
Triazene, Extracellular
Matrix Solution
Matrigel
®
Extracellular Matrix
Solution
Patz et al., 2005;
Doraiswamy et al., 2006b
Ovine endothelial cells
Gold
PEG scaffold
Ovsianikov et al., 2010
Ovine vascular smooth
muscle cells (vSMC)
Gold
PEG scaffold
Ovsianikov et al., 2010
Human keratinocytes
Gold, alginate
Matrigel
®
Koch et al., 2010
Human breast cancer
Alginate, gelatin
Gelatin, calcium chloride
Kingsley et al., 2013
Bovine Pulmonary Artery
Endothelial Cells (BPAEC)
Matrigel
®
Matrigel
®
Schiele et al., 2009
Rat neural stem cells
Matrigel
®
Matrigel
®
Schiele et al., 2009
Mouse myoblast
Matrigel
®
Matrigel
®
Schiele et al., 2009
because patterning proteins restricts cell migration to adhesion islands. Unrestricted cell growth following
LDW has been demonstrated with a Matrigel
®
coating on the substrate in earlier work (
Wu et al., 2003;
Schiele et al., 2009
), and more recently with a gelatin substrate coating (
Schiele et al., 2011
). An advan-
tage of the gelatin substrate is that the gelatin coating is temporary (as mentioned in Section 7.2.3), and
does not provide cells with a permanent scaffold or a large assortment of unknown complex signaling
factors. Thus, after the gelatin has liquefied during incubation, and is removed with the first media ex-
change, the cells remain attached to the underlying substrate. This allows various substrates and materials
to be utilized, provided the thin temporary gelatin layer is applied to enable viable transfers. Despite the
temporary nature of the gelatin layer, cells printed to the substrate maintain excellent pattern registry,
moving on average, less than 6
m
m from their initial location within half an hour of printing (
Schiele
et al., 2011
). One restriction of this technique is the limitation to a flat surface parallel to the ribbon, but
otherwise potentially any cell-adhesive material can be used with LDW, such as hydrogels or scaffold
materials with engineered topography.
Although LDW can potentially be used to study the effects of cell location and cell-cell interaction,
exploration of cellular phenomena after LDW patterning has been examined in only a few instances.
One study of embryonic stem cell (ESC) behavior examined the clustering of ESCs after LDW. ESCs
are pluripotent, meaning they can differentiate into any of the three primitive germ layers. It was shown
that following LDW, mouse ESCs (mESCs) maintained their pluripotency, as evidenced by their abil-
ity to express markers of all three primitive germ layers (
Raof et al., 2011
). An additional indication
of pluripotency is the formation of 3D cell clusters called embryoid bodies (EBs), which contain cells
of all three germ layers. The cell-cell interactions within the EB are greatly important. The size of the
EB, which may impart complex signals to cells within the structure, can influence differentiation, as
can the size of a stem cell colony (
Peerani et al., 2007
;
Bauwens et al., 2008
;
Lee et al., 2009b
). The
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